Molecular mechanism for change in permeability in brain microvascular endothelial cells induced by LPS
10.3969/j.issn.1672-7347.2010.11.003
- VernacularTitle:LPS诱导小鼠脑微血管内皮细胞通透性增高的分子机制
- Author:
Fang HE
;
Fei YIN
;
Jing PENG
;
Xiaolu DENG
;
Liwen WU
;
Ciliu ZHANG
- Publication Type:Journal Article
- Keywords:
LPS;
RhoA;
NF-κB;
F-actin;
brain microvascular endothelial cell
- From:
Journal of Central South University(Medical Sciences)
2010;35(11):1129-1137
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the molecular mechanism for change in permeability in brain microvascular endothelial cells (bEnd.3) induced by lipopolysaccharide (LPS). Methods Monolayers of bEnd.3 were exposed to LPS,in the presence or absence of exoenzyme C3 transferase. We monitored the monolayer barrier integrity by transendothelial electrical resistance assay (TEER),activity of RhoA by pull down assay,NF-κB by luciferase reporter assay,and F-actin dynamic structure by Rhodamine-phalloidin staining. Results Incubation of monolayers with LPS caused substantial barrier hyperpermeability. Under the had been treated for 3 and 12 h with LPS (P＜0.05). Such effects could be inhibited partly by pretreatment of RhoA inhibitor exoenzyme C3 transferase. LPS activated RhoA and NF-κB at 0.5 h. The C3 transferase could significantly reverse the NF-κB activation (P＜0.05). The F-actin rearrangments displayed in a time-dependent manner and occurred originally after the stimulation of LPS for 3 h,which could be diluted by the pretreatment of C3 transferase as well. Conclusion LPS induces the disruption of F-actin cytoskeleton and brain microvascular endothelial barrier integrity,in part,through RhoA and NF-κB activation. The mechanism underlying this pathophysiological effect of RhoA is to influence the disruption of the F-actin cytoskeleton by regulating NF-κB activites.
